To fulfill the comprehensive utilization of cellulose and hemicellulose components in bagasse for bacterial cellulose (BC) production, both bagasse acid and enzymatic hydrolysates were used for BC production by Gluconacetobacter xylinus. Although the BC accumulation rate was slower during the early period of fermentation in the bagasse acid hydrolysate than in the enzymatic hydrolysate, the highest BC yield (1.09 vs. 0.42g/L) was higher in the bagasse acid hydrolysate. The substrate utilization was evaluated in both bagasse acid and enzymatic hydrolysates, and glucose, xylose, and acetic acid were better carbon sources than arabinose and cellobiose for G. xylinus. The structure of the BC samples obtained from bagasse acid and enzymatic hydrolysates, including the microscopic morphology, functional groups, and crystals, was similar especially in the later phase of fermentation, which was analyzed by field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. Thus, both bagasse acid and enzymatic hydrolysates could be promising substrates for BC production. (c) 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017, 134, 45066.